ELISA and western blotting for the detection of Hsp70 and Hsp83 antigens of Leishmania donovani.

Jaspreet Kaur, Sukhbir Kaur
Author Information
  1. Jaspreet Kaur: Parasitology Laboratory, Department of Zoology, Panjab University, Chandigarh, 160014 India.
  2. Sukhbir Kaur: Parasitology Laboratory, Department of Zoology, Panjab University, Chandigarh, 160014 India.

Abstract

Visceral leishmaniasis (VL) is endemic in the tropical, sub-tropical regions of Asia, Africa, the Mediterranean, Southern Europe and South and Central America. Approximately 500,000 new cases are reported annually. Classically the diagnosis of VL is confirmed by the demonstration of the parasite in aspirates of spleen, bone marrow or liver which can yield false negative results, also these methods are invasive. In this study, we aimed to evaluate the serodiagnostic potential of two heat shock proteins (Hsps) i.e. Hsp70 and Hsp83 in combination by ELISA and western blotting in mouse model. Parasite proteins were separated by SDS-PAGE and transferred to PVDF membranes which were further incubated with serum samples of infected mice for visualizing different bands. Both the bands i.e. Hsp70 and Hsp83 were simultaneously visualized in all infected groups on different post infection days. The presence of both the antigens was also detected by sandwich ELISA. The results suggest that the simultaneous occurrence of both the antigens Hsp70 and Hsp83 may be useful in serodiagnostic assay of VL as it may reduce the need for traumatic bone marrow sampling and risky spleen aspiration.

Keywords

References

  1. Trans R Soc Trop Med Hyg. 1995 Mar-Apr;89(2):159-62 [PMID: 7778140]
  2. Trop Med Int Health. 2004 Jun;9(6):724-9 [PMID: 15189464]
  3. J Clin Microbiol. 1991 Jul;29(7):1429-35 [PMID: 1885738]
  4. Clin Vaccine Immunol. 2006 Feb;13(2):271-6 [PMID: 16467337]
  5. Am J Trop Med Hyg. 2005 Aug;73(2):269-71 [PMID: 16103587]
  6. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):775-9 [PMID: 8421715]
  7. J Gen Virol. 1976 Oct;33(1):165-7 [PMID: 789817]
  8. J Egypt Soc Parasitol. 2008 Dec;38(3):1027-36 [PMID: 19209783]
  9. Ann Trop Med Parasitol. 1978 Jun;72(3):213-18 [PMID: 666392]
  10. Acta Trop. 2002 Jun;82(3):339-48 [PMID: 12039673]
  11. Immunol Lett. 1995 Dec;48(3):209-14 [PMID: 8867853]
  12. J Clin Microbiol. 2002 Feb;40(2):475-9 [PMID: 11825959]
  13. Trop Med Int Health. 2007 Feb;12(2):284-9 [PMID: 17300637]
  14. Nature. 1970 Aug 15;227(5259):680-5 [PMID: 5432063]
  15. Infect Immun. 1997 Dec;65(12):5368-70 [PMID: 9393843]
  16. Parasitol Today. 1992 Mar;8(3):104-5 [PMID: 15463585]
  17. Parasite Immunol. 2011 Feb;33(2):95-103 [PMID: 21226722]
  18. J Immunol. 1990 May 15;144(10):3987-91 [PMID: 2110215]
  19. Clin Diagn Lab Immunol. 2002 Sep;9(5):951-8 [PMID: 12204943]
  20. Exp Parasitol. 1997 Mar;85(3):249-63 [PMID: 9085922]
  21. Acta Trop. 2001 Jan 15;78(1):11-6 [PMID: 11164745]
  22. J Immunol. 1990 Jan 15;144(2):699-706 [PMID: 2295807]
  23. APMIS. 1993 Aug;101(8):642-6 [PMID: 8105816]
  24. Immunol Today. 1991 Mar;12(3):A38-41 [PMID: 2069677]
  25. Clin Exp Immunol. 1996 Dec;106(3):454-61 [PMID: 8973612]
  26. Annu Rev Immunol. 1990;8:401-20 [PMID: 2188668]
  27. Infect Immun. 2004 Feb;72(2):863-70 [PMID: 14742530]
  28. Trans R Soc Trop Med Hyg. 2004 Feb;98(2):102-10 [PMID: 14964810]
  29. Br J Biomed Sci. 2002;59(4):218-22 [PMID: 12572956]
  30. J Infect Dis. 1996 Mar;173(3):758-61 [PMID: 8627048]
  31. Crit Rev Microbiol. 1995;21(2):123-52 [PMID: 7639932]
  32. Immunol Today. 1990 Apr;11(4):129-36 [PMID: 2187470]
  33. Cell. 1989 Oct 6;59(1):5-8 [PMID: 2676194]
  34. J Trop Med Hyg. 1994 Dec;97(6):333-40 [PMID: 7966534]
  35. Am J Trop Med Hyg. 1995 May;52(5):427-8 [PMID: 7539596]
  36. BMJ. 2006 Oct 7;333(7571):723 [PMID: 16882683]
  37. Acta Trop. 2011 Jul;119(1):50-6 [PMID: 21530477]
  38. Acta Trop. 1996 Sep;62(1):45-56 [PMID: 8971277]
  39. J Infect Dis. 1988 Jun;157(6):1212-20 [PMID: 3373022]
  40. J Biol Chem. 1951 Nov;193(1):265-75 [PMID: 14907713]
  41. Trans R Soc Trop Med Hyg. 1992 Sep-Oct;86(5):505-7 [PMID: 1475815]

Word Cloud

Created with Highcharts 10.0.0Hsp70Hsp83ELISAVLblottingantigensspleenbonemarrowresultsalsoserodiagnosticproteinsiewesterninfecteddifferentbandsmayVisceralleishmaniasisendemictropicalsub-tropicalregionsAsiaAfricaMediterraneanSouthernEuropeSouthCentralAmericaApproximately500000newcasesreportedannuallyClassicallydiagnosisconfirmeddemonstrationparasiteaspirateslivercanyieldfalsenegativemethodsinvasivestudyaimedevaluatepotentialtwoheatshockHspscombinationmousemodelParasiteseparatedSDS-PAGEtransferredPVDFmembranesincubatedserumsamplesmicevisualizingsimultaneouslyvisualizedgroupspostinfectiondayspresencedetectedsandwichsuggestsimultaneousoccurrenceusefulassayreduceneedtraumaticsamplingriskyaspirationdetectionLeishmaniadonovaniWestern

Similar Articles

Cited By